Gdh kit

Glutamate dehydrogenase (GDH), a mitochondrial enzyme, plays an important role in controlling insulin secretion. GDH activity was measured using the GDH kit (Abcam, Cambridge, UK) as per the vendor’s protocol. Briefly, the mitochondrial samples from the pancreas and heart of control Wistar and GK diabetic rats were incubated in a reaction buffer, containing glutamate and the NADH produced was measured spectrophotometrically using the UV-1800 spectrophotometer (Schimadzu Corporation, Kyoto, Japan) at 450 nm using a standard curve.

Activity of hexokinase, the rate-limiting enzyme in the glycolytic pathway, was measured using a hexokinase colorimetric assay kit (Sigma-Aldrich, St. Louis, MO, USA) as per the vendor’s protocol. Briefly, it is a coupled enzyme assay, resulting in the production of NADH, which reduces a colorless probe, producing a colorimetric (450 nm) product proportional to the hexokinase activity.
Activity of glutamate dehydrogenase (GDH), a mitochondrial enzyme, was measured using a GDH kit (Abcam, Cambridge, UK) as per the manufacturer’s instructions. Briefly, mitochondria from the different tissues were treated with a reaction buffer, containing glutamate as a specific substrate and the NADH generated was determined spectrophotometrically using the UV-1800 spectrophotometer (Schimadzu Corporation, Kyoto, Japan) at 450 nm.